Specification
Sheet
Please note
Some information
is Lot dependent,
Monoclonal Mouse
Anti-Human CD62P,P-Selectin,
Clone 1E3
Code No,M 7199
Lot 038,Edition 06.12.00
Antigen synonyms Platelet activation-dependent granule membrane protein (PADGEM) or GMP-140,
Introduction The CD62P antigen is a 140 kDa transmembrane glycoprotein present on
megakaryocytes,activated platelets and activated endothelial cells,In unstimulated
platelets and endothelial cells CD62P is stored in the membranes of secretory granules
and shows no surface immunostaining with anti-CD62P,When platelets and endothelial
cells are activated by agonists such as thrombin,the granules rapidly fuse with the plasma
membrane,which will express CD62P on the surface (1-3),CD62P is a receptor for
neutrophils and monocytes when it is expressed on activated platelets and endothelium
(4),
CD62P is a member of the small selectin family of cellular adhesion molecules which also
includes CD62E (E-selectin) and CD62L (L-selectin) (5,6),Its structure,similar to the
other members of the selectin family,consists of an N-terminal lectin-like domain of C-
type (6) followed by an epidermal growth factor-like domain,nine short consensus repeats
(SCR) related to those in complement-binding proteins,a transmembrane region and a
short cytoplasmic tail which bears no homology to the CD62E and CD62L cytoplasmic
domains (3),The major CD62P ligand on neutrophils is the P-Selectin Glycoprotein
Ligand 1,CD162 (PSGL-1) (5,7),
Presentation Monoclonal mouse antibody supplied in liquid form as tissue culture supernatant (RPMI
1640 medium containing fetal calf serum) dialysed against 0.05 mol/L Tris/HCl,pH 7.2
containing 15 mmol/L NaN
3
,
Mouse Ig concentration,170 mg/L,
Isotype,IgG2a,kappa,
Total protein concentration,11.7 g/L,
Storage 2-8 °C,
Clone 1E3,(8),
Immunogen Fixed thrombin-activated human platelets,
Specificity/reactivity Anti-CD62P,1E3,was included in the Sixth International Workshop and Conference on
Human Leucocyte Differentiation Antigens (Kobe 1996),and studies by a number of
laboratories confirmed its reactivity with the CD62P antigen (9),
The antibody stains megakaryocytes,activated platelets and activated endothelium,Other
normal cells do not react with the antibody,
Anti-CD62P,1E3,is useful for assessing platelet activation in acute coronary syndromes
by flow cytometry (10),
(2)
- 2 -
Staining procedures Formalin-fixed and paraffin-embedded sections
Can be used on formalin-fixed,paraffin-embedded tissue sections,Antigen retrieval,such as
by heating in DAKO Target Retrieval Solution,code No,S 1700 is mandatory,The slides
should not be allowed to dry out during this treatment or during the following
immunohistochemical staining procedure,
For tissue sections sensitive staining techniques are recommended,such as the LSAB?+ or
the EnVision?+ system,
The antibody may be used at a dilution of 1:25-1:50 with the EnVision?+ system when
tested on formalin-fixed,paraffin-embedded sections of human tonsil,
Frozen sections and cell smears
Can be used for labelling acetone-fixed cryostat sections or fixed cell smears,The antibody
may be used at a dilution of 1:25-1:50 in the APAAP technique when tested on acetone-fixed
sections,
This is a guideline only; an optimal dilution should be determined by the individual laboratory,
References (1) McEver RP,Martin MN,A monoclonal antibody to a membrane glycoprotein binds only to activated
platelets,J Biol Chem 1984;259:9799-804,
(2) Stenberg PE,McEver RP,Shuman MA,Jacques YV,Bainton DF,A platelet alpha-granule membrane
protein (GMP-140) is expressed on the plasma membrane after activation,J Cell Biol 1985;101:880-6,
(3) Johnston GI,Cook RG,McEver RP,Cloning of GMP-140,a granule membane protein of platelets and
endothelium,sequence similarity to proteins involved in cell adhesion and inflammation,Cell
1989;56:1033-44,
(4) McEver RP,GMP-140,a receptor for neutrophils and monocytes on activated platelets and endothelium,
J Cell Biochem 1991;45:156-61,
(5) McEver RP,Moore KL,Cummings RD,Leukocyte trafficking mediated by selectin-carbohydrate
interactions,J Biol Chem 1995;270:11025-8,
(6) Lasky LA,Selectin-carbohydrate interactions and the initiation of the inflammatory response,Annu Rev
Biochem 1995;64:113-39,
(7) Rosen SD,Bertozzi CR,Two selectins converge on sulphate,Leukocyte adhesion,Curr Biol 1996;6:261-
4,
(8) Carmody MW,Ault KA,Mitchell JG,Rote NS,Ah-Kau NG,Production of monoclonal antibodies specific
for platelet activation antigens and their use in evaluating platelet function,Hybridoma 1990;9:631-41,
(9) de Haas M,von dem Borne AEG,PL8,CD62P workshop panel report,In,Kishimoto T et al.,eds,
Leucocyte Typing VI,White Cell Differentiation Antigens,New York - London,Garland Publishing,Inc.,
1997:663-4,
(10) Becker RC,Tracy RP,Bovill EG,Mann KG,Ault K,The clinical use of flow cytometry for assessing
platelet activation in acute coronary syndromes,TIMI-III Thrombosis and Anticoagulation Group,Coron
Artery Dis 1994;5:339-45,
M 7199/TJU/06.12.00